Molecular cloning, overexpression and characterization of the raw-starch-digesting α-amylase of Bacillus amyloliquefaciens

被引：6

作者：

Gangadharan, Dhanya ^{[1
]}

Ramachandran, Priya ^{[2
]}

Paramasamy, Gunasekaran ^{[2
]}

Pandey, Ashok ^{[1
]}

Nampoothiri, K. Madhavan ^{[1
]}

机构：

[1] CSIR, NIIST, Div Biotechnol, Trivandrum 695019, Kerala, India

[2] Madurai Kamaraj Univ, Sch Biol Sci, Ctr Excellence Genom Sci, Dept Genet, Madurai 625021, Tamil Nadu, India

来源：

BIOLOGIA | 2010年 / 65卷 / 03期

关键词：

Bacillus amyloliquefaciens; alpha-amylase; cloning; overexpression; raw starch digestion; BIOCHEMICAL-CHARACTERIZATION; NUCLEOTIDE-SEQUENCE; BINDING DOMAIN; BETA-AMYLASE; GLUCOAMYLASE; GENE; PURIFICATION; EXPRESSION; DEGRADATION; PROMOTER;

D O I：

10.2478/s11756-010-0042-6

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Raw starch is the most abundant source of glucose in the world. Therefore, finding enzymes capable of digesting raw starch would find high industrial demand. The alpha-amylase gene of Bacillus amyloliquefaciens ATCC 23842 was amplified, cloned and overexpressed in Escherichia coli BL21 (DE3) cells. The recombinant enzyme was purified to apparent homogeneity using ion exchange and gel filtration chromatography. The raw-starch digestibility of the purified enzyme was characterized by studying the hydrolysis and adsorption rate on a variety of raw starches (potato, cassava, corn, wheat and rice). The raw-starch digestion was further confirmed by scanning electron microscopy studies, which revealed an effective rate of hydrolysis. The kinetic studies revealed a relatively low K (m) of 2.76 mg/mL, exhibiting high affinity towards the soluble starch as the most preferred substrate and the inhibition kinetic studies revealed a high K (i) value (350 mM).

引用

页码：392 / 398

页数：7

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